OBJECTIVE:To determine the content of gallic acid in viviparous bistort rhizome by HPLC.METHODS:The chromatographic column was Nova-Pak C 18 and the mobile phase was0.2%methanol methyl cyanide solution-0.1%phos?phoric acid-0.15%triethylamin(3∶50∶47),the flow rate was1.0ml/min and the detection wavelength was271nm.RE?SULTS:It took a good linear relationship when the sample size of gallic acid was within the range of0.028～0.14?g(r=0.9991),the average recovery was99.05%(RSD=4.0%).CONCLUSION:The method is convenient,fast,reproducible,which can be used for the content determination of gallic acid in viviparous bistort rhizome.

Objective To study and observe the engraftment of donor cells from unrelated cord blood into adult patients with severe aplastic anemia (SAA) and the outcome of allo-CBSCT. Methods Six patients received cord blood provided by Guangzhou Cord Blood Bank, for three of which one unit of cord blood was given in a procedure, whereas for other 3 patients, 2 units of cord blood were infused at the same time in a transplant protocol. In all 9 units of umbilical cord blood (UCB) infused, UCB units contained 1.6 ～ 10.7 nucleated cells/kg body weight of the recipient after thawing. The patients were conditioned with decreased dosage of immunosuppressive agents of CTX (60?mg/kg) and ALG (120?mg/kg). The engraftment state of the donor cells into recipients was confirmed by microsatellite DNA fingerprinting and fluorescent quantitative PCR analysis. Results Engrafted evidence has been found in 5 patients by molecular biology analyses showing donor-recipient mixed chimerism post transplant which was stable and persistent. After a median follow-up of 21 months (range 7～50), 4 patients were alive and disease free. One patient died of severe infection in the 3rd month from transplant, though the evidence of engraftment was obvious. Another patient also died in the early stage posttransplant of serious aspergillus infection without the engrafted proof.Conclusion Durable donor-recipient stable mixed chimerism can be achieved by unrelated UCBT in patients with SAA. Umbilical cord blood could be employed as a source of hematopoietic stem cell for adult transplantation.

Objective To explore the possibility of cytochrome P-450 3A5 (CYP3A5) genotype as a major factor to guide individualized employment of cyclosporin A(CsA) through a comparative study of CsA concentrations in the peripheral blood of hemopoietic stem cell transplant recipients with different CYP3A5 genotypes. Methods Olymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to genotype CYP3A5 gene, and CsA concentrations were detected by a commercial fluorescence polarization immunoassay. Result There were significant differences in CsA concentrations, including standardized trough concentrations C0 and two h peak concentrations C2 , in the two CYP3A5 genotypes found in our samples, and both C0 and C2 in wild homozygotes were lower than heterozygotes. Conclusion CYP3A5 polymorphism is highly associated with CsA concentrations in hemopoietic stem cell transplant recipients, and CYP3A5 genotype may be a predictor to the dose requirement before clinically employment of CsA.

Objective To investigate the antimicrobial properties of human amniotic homogenate supernatant (HAHS) in order to found a theoretical base for the use of human amniotic membrane in ophthalmological field. MethodsFresh human amniotic membranes were used to make HAHS and acellular amniotic membranes. Then, we observed their antimicrobial effects and antimicrobial spectrums, compared the antimicrobial capacity with 10 commonly used antibiotics in eyes, and investigated the effects of time, temperature and pH value on the antimicrobial capacity. Finally, transmission electron microscopy (TEM) was used to explore the possible targeting site of the antibiosis. ResultsHuman amnion membrane contained antimicrobial components locating in its epithelial cells. HAHS had a broad antimicrobial spectrum and was steady in nature. Its antimicrobial capacity was stronger than those of sulfasulfonamide, chloromycetin and cefuroxime sodium. TEM indicated that the antimicrobial effect were exerted through plasma membrane of microorganism. ConclusionHAHS can be an effective and convenient treatment for ocular surface infectious diseases. Traditional amnion transplantation should employ fresh human amniotic membranes containing complete epithelial lamina to reconstruct the ocular surface.

Objective Pulmonary benign metastasizing leiomyoma (PBML) is a rare entity that leiomyoma of uterus metastasized to the lung.The clinical characteristics of this rare disease were analyzed in this article.Methods The detailed clinical records of 7 patients diagnosed as PBML at Peking Union Medical College Hospital between January 2001 and June 2015 were reviewed.Results All patients were women with median age of 44 years (range 28-62).Symptoms included dyspnea (2/7),chest pain (1/7),cyanosis (1/7),cough (1/7) and bloody sputum (1/7),while 4/7 cases were asymptomatic.Six patients had the past-history of leiomyoma of uterus 20 months to 14 years ago among whom 5 patients received hysterectomy.Chest CT showed bilateral,random-distributed multiple round solid nodules,or diffuse-distributed miliary nodules,or single solid nodule,even some small cavities.Extra-pulmonary metastasis was found in left superclavicular lymph node (1 case) and right heart (1 case).Histological tissues were obtained by video-assisted thoracic surgery lung biopsy (4/7),mass resection on tricuspid valve (1/7),transbronchil lung biopsy (1/7),and CT-guided percutaneous lung biopsy (1/7).Pathology showed an interlacing pattern by spindle cells having elongated nuclei without cellular atypia.Ki-67 index was less than 1％.Molecules such as smooth muscle antibody,estrogen receptor (ER) and progestrone receptor (PR) were positive in immunohistochemistry staining.Neither letrozole nor zoladex was effective.Two patients responded to bilateral adnexectomy,presenting as shrunk nodules.No relapsed disease was seen in one patient with single nodule after resection.There was only one patient with disease-related mortality,whose chest CT showed milliary nodules.Conclusion Although CT findings of PBML are similar to malignancies,the clinical outcome is good.Despite the positive expression of ER and PR,the effectiveness of hormone related treatment is limited.And periodical follow up is suggested even to those uneventful patients.

A new method for noninvasive prenatal diagnosis of fetal sex was developed by using single-cell PEP-PCR techniques. Micromamipulation techniques were used to obtain single fetal cells from 273 maternal blood samples. The genome of single cells was preamplified by PEP and SRY genes were analyzed by PCR method. The SRY genes of 149 samples were detected by the new method among 153 samples carrying male fetus, while 119 out of 120 samples carrying female fetus were proved negative for SRY genes. The sensitivity and specificity of the new method were 97.39% and 99.17% respectively and the correct rate was 98.17%. The new method has the advantage of high sensitivity and specificity in noninvasive prenatal diagnosis of fetal sex and provides the basis of other researches such as sex-linked inherited diseases.
Chromosomes, Human, Y ; Erythroblasts/chemistry ; Fetus/*cytology ; Genes, sry/genetics ; Genetic Diseases, Inborn/diagnosis ; Maternal-Fetal Exchange/genetics ; Polymerase Chain Reaction/methods ; Pregnancy/*blood ; Prenatal Diagnosis/*methods ; *Sex Determination (Genetics)

Objective To evaluate the safety and efficacy of icotinib as first-line therapy in Chinese non-small cell lung cancer (NSCLC) patients harboring epidermal growth factor receptor (EGFR) sensitive mutations.Methods Patients with stage ⅢB/Ⅳ NSCLC who had EGFR sensitive mutation and had no previous treatment were enrolled into this study.The response rates , progress free survival ( PFS) , overall survival ( OS ) , and the safety were analyzed.Results Ninety advanced adenocarcinoma patients were enrolled in this study , 44 patients had partial response ( PR ) , 42 patients had stable disease ( SD ) , 4 patients had progressive disease (PD), with an overall response rate (ORR) of 48.9%, and a disease control rate (DCR) of 95.6%.The median PFS was 14.9 months (95%CI 13.5-16.3) and the OS was 37.0 weeks ( 95%CI 27.9 -46.1 ).Patients with brain metastases showed higher ORR ( P =0.049 ).Patients with stage ⅢB had longer PFS than those with stage Ⅳ( P=0.007 ).The most common adverse events were grade 1 -2 skin rash (38 patients, 40.9%).Other adverse events included dry skin , oral mucositis, diarrhea and liver function injury.Three patients withdrew because of severe liver injury or skin rash.No treatment related mortality occurred .Conclusions Icotinib is effective and safe as first-line treatment for Chinese advanced NSCLC patients with EGFR sensitive mutation.

Objective To observe the effect of postoperative central vault on the anterior chamber after implantation of im-plantable contact lens V4c .Methods To carry out a retrospective study of 56 eyes with implantable contact lens V4c implantation in Chongqing Bright Eye Hospital from November 2 ,2014 to February 5 ,2015 ,examined anterior chamber angle before surgery ,the anterior chamber angular width (ACA) and anterior chamber volume (ACV) were measured by pentacam .the postoperative central vault and ACA and ACV during the first and third and sixth months after surgery were measured ,the pigment grade in anterior chamber after operation were recorded .The change of ACA and ACV of the 56 eyes was correlation analyzed .Results At first month after surgery ,the postoperative central vault was 250 ~ 765 μm ,the reduced amount of ACA was (13 .67 ± 1 .67)° ,the re-duced amount of ACV was (94 .23 ± 17 .02)mm3 .At the third month ,the postoperative central vault of the 55 eyes was 239 ~ 761μm ,the reduced amount of ACA was (13 .70 ± 1 .68)° ,The reduced amount of ACV was (93 .6 ± 17 .48)mm3 .At the sixth month , the postoperative central vault of the 51 eyes was 246 ~ 761 μm ,the reduced amount of ACA was (13 .60 ± 1 .69)° ,the reduced a-mount of ACV was(94 .29 ± 17 .80)mm3 .No case of pigment was found for grade Ⅱ .The statistical comparison of the amount of ACV and ACA decreased in different postoperative central vaults showed that the difference was statistically significant ,there were statistically significant differences in the amount of ACV and ACA between different central postoperative central vault(P< 0 .05) . Conclusion Anterior chamber angle was narrowed and anterior chamber volume decreased in the posterior chamber intraocular lensimplant-ation after implantable contact lens V4c implantation ,the postoperative central vault is related to the change of anterior chamber angle and ACV .Appropriate postoperative central vault has positive clinical significance for keeping anterior chamber angle open .

Objective To identify and quantitate the conjunctival aerobic bacterial flora from healthy people of 3 age groups in Chongqing city.Methods Samples taken with moist swabs from the lower fornix of both eyes of 33 children(9.85?0.57 years old),31 youths(21.23?0.88 years old)and 30 elders(72.97?4.94 years old)were given bacterial cultures.Positive ones were identified and colony forming units(CFUs)were counted.Bacterial quantity was expressed as the number of CFUs divided by the weight of each sample measured before and after collection.Results The elders showed a higher isolation frequency(93.33%)than children(45.45%)and youths(50.00%)which had statistically difference(P

OBJECTIVE To type the genes of plasmid DNA in 54 clinical Klebsiella pneumoniae isolates producing extended -spectrum beta-lactamases (SHV) by denaturing high-performance liquid chromatography (DHPLc) and evaluate their sensitivity and specificity, and explore a rapid and convenient method for detecting the antibiotic resistance of K. pneumoniae. METHODS Plasmid DNA from each extended-spectrum beta-lactamase (SHV) producing strain was subjected to PCR amplification. After we performed DNA sequencing of these amplicons and identification of mutation and their genotype, DHPLc was undertaken to investigate whether its results correlate the distinctive chromatogram with each genotype. RESULTS All the strains were found abnormal elution peaks (two or three peaks) which were different from each other. The result of DNA sequencing demonstrated that all the strains had DNA mutation in comparison with SHV-1. Moreover, DHPLc could produce specific peak patterns that correlate with genotype. CONCLUSIONS The sensitivity of DHPLc is 100% in this study. And each genotype is corresponded to specific peak pattern. So we can use DHPLc technique to type the genes of plasmid DNA in K. pneumoniae and detect mutations rapidly. DHPLc not only has high accuracy , but also is a convenient and rapid technique for the detection of mutation in the bacterial genome. It has a great potential clinical value.